Methotrexate-Induced Decrease in Embryonic 5-Methyl-Tetrahydrofolate Is Irreversible with Leucovorin Supplementation

Tseng Ting Kao, Gang Hui Lee, Chi Chang Fu, Bing Hung Chen, Li Ting Chen, Tzu Fun Fu

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Folate is a nutrient crucial for rapidly growing tissues, including developing embryos and cancer cells. Folate participates in the biosynthesis of nucleic acids, proteins, amino acids, S-adenosylmethionine, many neurotransmitters, and some vitamins. The intracellular folate pool consists of different folate adducts, which carry one-carbon units at three different oxidative states and participate in distinct biochemical reactions. Therefore, the content and dynamics of folate adducts will affect the homeostasis of the metabolites generated in these folate-mediated reactions. Currently, the knowledge on the level of each individual folate adduct in developing embryos is limited. With an improved high-performance liquid chromatography protocol, we found that tetrahydrofolate (THF), the backbone of one-carbon carrier, gradually increased and became dominant in developing zebrafish embryos. 5-methyl-tetrahydrofolate (5-CH3-THF) was abundant in unfertilized eggs but decreased rapidly when embryos started to proliferate and differentiate. 10-formyltetrahydrofolate at first increased after fertilization, and then dropped dramatically before reaching a sustained level at later stages. Dihydrofolate (DHF) slightly decreased initially and remained low throughout embryogenesis. Exposure to methotrexate significantly decreased 5-CH3-THF levels and increased DHF pools, besides causing brain ventricle anomaly. Rescuing with leucovorin partly reversed the abnormal phenotype. Unexpectedly, the level of 5-CH3-THF remained low even when leucovorin was added for rescue. Our results show that different folate adducts fluctuated significantly and differentially in concert with the physiological requirement specific for the corresponding developmental stages. Furthermore, methotrexate lowered the level of 5-CH3-THF in developing embryos, which could not be reversed with folate supplementation and might be more substantial to cellular methylation potential and epigenetic control than to nucleotide synthesis.

Original languageEnglish
Pages (from-to)326-337
Number of pages12
JournalZebrafish
Volume10
Issue number3
DOIs
Publication statusPublished - 2013 Sep 1

Fingerprint

methotrexate
Leucovorin
Folic Acid
Methotrexate
folic acid
embryo (animal)
Embryonic Structures
Carbon
5,6,7,8-tetrahydrofolic acid
tetrahydrofolic acid
S-Adenosylmethionine
S-adenosylmethionine
carbon
Zebrafish
chemical reactions
neurotransmitters
Danio rerio
Fertilization
Epigenomics
epigenetics

All Science Journal Classification (ASJC) codes

  • Animal Science and Zoology
  • Developmental Biology

Cite this

Kao, Tseng Ting ; Lee, Gang Hui ; Fu, Chi Chang ; Chen, Bing Hung ; Chen, Li Ting ; Fu, Tzu Fun. / Methotrexate-Induced Decrease in Embryonic 5-Methyl-Tetrahydrofolate Is Irreversible with Leucovorin Supplementation. In: Zebrafish. 2013 ; Vol. 10, No. 3. pp. 326-337.
@article{0c8d2345eb1a41f5a8ab039e5c48f7aa,
title = "Methotrexate-Induced Decrease in Embryonic 5-Methyl-Tetrahydrofolate Is Irreversible with Leucovorin Supplementation",
abstract = "Folate is a nutrient crucial for rapidly growing tissues, including developing embryos and cancer cells. Folate participates in the biosynthesis of nucleic acids, proteins, amino acids, S-adenosylmethionine, many neurotransmitters, and some vitamins. The intracellular folate pool consists of different folate adducts, which carry one-carbon units at three different oxidative states and participate in distinct biochemical reactions. Therefore, the content and dynamics of folate adducts will affect the homeostasis of the metabolites generated in these folate-mediated reactions. Currently, the knowledge on the level of each individual folate adduct in developing embryos is limited. With an improved high-performance liquid chromatography protocol, we found that tetrahydrofolate (THF), the backbone of one-carbon carrier, gradually increased and became dominant in developing zebrafish embryos. 5-methyl-tetrahydrofolate (5-CH3-THF) was abundant in unfertilized eggs but decreased rapidly when embryos started to proliferate and differentiate. 10-formyltetrahydrofolate at first increased after fertilization, and then dropped dramatically before reaching a sustained level at later stages. Dihydrofolate (DHF) slightly decreased initially and remained low throughout embryogenesis. Exposure to methotrexate significantly decreased 5-CH3-THF levels and increased DHF pools, besides causing brain ventricle anomaly. Rescuing with leucovorin partly reversed the abnormal phenotype. Unexpectedly, the level of 5-CH3-THF remained low even when leucovorin was added for rescue. Our results show that different folate adducts fluctuated significantly and differentially in concert with the physiological requirement specific for the corresponding developmental stages. Furthermore, methotrexate lowered the level of 5-CH3-THF in developing embryos, which could not be reversed with folate supplementation and might be more substantial to cellular methylation potential and epigenetic control than to nucleotide synthesis.",
author = "Kao, {Tseng Ting} and Lee, {Gang Hui} and Fu, {Chi Chang} and Chen, {Bing Hung} and Chen, {Li Ting} and Fu, {Tzu Fun}",
year = "2013",
month = "9",
day = "1",
doi = "10.1089/zeb.2013.0876",
language = "English",
volume = "10",
pages = "326--337",
journal = "Zebrafish",
issn = "1545-8547",
publisher = "Mary Ann Liebert Inc.",
number = "3",

}

Methotrexate-Induced Decrease in Embryonic 5-Methyl-Tetrahydrofolate Is Irreversible with Leucovorin Supplementation. / Kao, Tseng Ting; Lee, Gang Hui; Fu, Chi Chang; Chen, Bing Hung; Chen, Li Ting; Fu, Tzu Fun.

In: Zebrafish, Vol. 10, No. 3, 01.09.2013, p. 326-337.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Methotrexate-Induced Decrease in Embryonic 5-Methyl-Tetrahydrofolate Is Irreversible with Leucovorin Supplementation

AU - Kao, Tseng Ting

AU - Lee, Gang Hui

AU - Fu, Chi Chang

AU - Chen, Bing Hung

AU - Chen, Li Ting

AU - Fu, Tzu Fun

PY - 2013/9/1

Y1 - 2013/9/1

N2 - Folate is a nutrient crucial for rapidly growing tissues, including developing embryos and cancer cells. Folate participates in the biosynthesis of nucleic acids, proteins, amino acids, S-adenosylmethionine, many neurotransmitters, and some vitamins. The intracellular folate pool consists of different folate adducts, which carry one-carbon units at three different oxidative states and participate in distinct biochemical reactions. Therefore, the content and dynamics of folate adducts will affect the homeostasis of the metabolites generated in these folate-mediated reactions. Currently, the knowledge on the level of each individual folate adduct in developing embryos is limited. With an improved high-performance liquid chromatography protocol, we found that tetrahydrofolate (THF), the backbone of one-carbon carrier, gradually increased and became dominant in developing zebrafish embryos. 5-methyl-tetrahydrofolate (5-CH3-THF) was abundant in unfertilized eggs but decreased rapidly when embryos started to proliferate and differentiate. 10-formyltetrahydrofolate at first increased after fertilization, and then dropped dramatically before reaching a sustained level at later stages. Dihydrofolate (DHF) slightly decreased initially and remained low throughout embryogenesis. Exposure to methotrexate significantly decreased 5-CH3-THF levels and increased DHF pools, besides causing brain ventricle anomaly. Rescuing with leucovorin partly reversed the abnormal phenotype. Unexpectedly, the level of 5-CH3-THF remained low even when leucovorin was added for rescue. Our results show that different folate adducts fluctuated significantly and differentially in concert with the physiological requirement specific for the corresponding developmental stages. Furthermore, methotrexate lowered the level of 5-CH3-THF in developing embryos, which could not be reversed with folate supplementation and might be more substantial to cellular methylation potential and epigenetic control than to nucleotide synthesis.

AB - Folate is a nutrient crucial for rapidly growing tissues, including developing embryos and cancer cells. Folate participates in the biosynthesis of nucleic acids, proteins, amino acids, S-adenosylmethionine, many neurotransmitters, and some vitamins. The intracellular folate pool consists of different folate adducts, which carry one-carbon units at three different oxidative states and participate in distinct biochemical reactions. Therefore, the content and dynamics of folate adducts will affect the homeostasis of the metabolites generated in these folate-mediated reactions. Currently, the knowledge on the level of each individual folate adduct in developing embryos is limited. With an improved high-performance liquid chromatography protocol, we found that tetrahydrofolate (THF), the backbone of one-carbon carrier, gradually increased and became dominant in developing zebrafish embryos. 5-methyl-tetrahydrofolate (5-CH3-THF) was abundant in unfertilized eggs but decreased rapidly when embryos started to proliferate and differentiate. 10-formyltetrahydrofolate at first increased after fertilization, and then dropped dramatically before reaching a sustained level at later stages. Dihydrofolate (DHF) slightly decreased initially and remained low throughout embryogenesis. Exposure to methotrexate significantly decreased 5-CH3-THF levels and increased DHF pools, besides causing brain ventricle anomaly. Rescuing with leucovorin partly reversed the abnormal phenotype. Unexpectedly, the level of 5-CH3-THF remained low even when leucovorin was added for rescue. Our results show that different folate adducts fluctuated significantly and differentially in concert with the physiological requirement specific for the corresponding developmental stages. Furthermore, methotrexate lowered the level of 5-CH3-THF in developing embryos, which could not be reversed with folate supplementation and might be more substantial to cellular methylation potential and epigenetic control than to nucleotide synthesis.

UR - http://www.scopus.com/inward/record.url?scp=84883403118&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84883403118&partnerID=8YFLogxK

U2 - 10.1089/zeb.2013.0876

DO - 10.1089/zeb.2013.0876

M3 - Article

C2 - 23758124

AN - SCOPUS:84883403118

VL - 10

SP - 326

EP - 337

JO - Zebrafish

JF - Zebrafish

SN - 1545-8547

IS - 3

ER -